Method for embedding a biological sample in a transparent matrix for analysis using single plane illumination microscopy

ABSTRACT

The invention is directed to method for positioning and aligning a preferably biological sample in the detection area of the objective of a microscope arrangement. According to the invention, the method mentioned above has the following method steps: a sample is introduced into a transparent medium, preferably agarose gel, which is initially liquid; the medium is changed from the liquid state to the solid state, wherein the sample is fixated within the medium, but the transparency of the medium is retained; the solidified medium is positioned in the microscope arrangement in such a way that the sample enclosed therein is situated in the detection area of the objective. Further, a device is proposed for positioning and aligning a preferably biological sample in the detection area of the objective of a microscope arrangement.

The present application claims priority from PCT Patent Application No.

PCT/EP2009/004081 filed on Jun. 6, 2009, which claims priority fromGerman Patent Application No. DE 10 2008 027 784.3 filed on Jun. 11,2008, the disclosure of which is incorporated herein by reference in itsentirety.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The invention is directed to a method and a device for positioning andaligning a preferably biological sample in the detection area of theobjective in a microscope arrangement.

The method according to the invention can be applied particularly inconnection with single plane illumination microscopy (SPIM). Thismicroscopy method is a special method of widefield microscopy by whichimage data for a three-dimensional image of the sample are obtained onthe basis of optical sections through different planes of the sample.

2. Description of Related Art

SPIM technology is described, for example, in Stelzer et al., OpticsLetter 31, 1477 (2006), Stelzer et al., Science 305, 1007 (2004), DE 10257 423 A1, and WO 2004/0530558 A1.

The viewing direction on the sample is changed repeatedly so that imagedata are not obtained from just one viewing direction on the sample.Every time the viewing direction is readjusted, the sample must bepositioned and aligned relative to the detection objective.

At present, the positioning and alignment of the sample to be examinedis carried out manually using commercially available equipment and istherefore extremely time-consuming and is accordingly only suitable forlaboratory analyses of individual samples.

However, particularly with regard to high volumes in industrialapplications, there is an increasing demand for detection of a largenumber of samples successively in time with the highest possiblethroughput per time unit.

SUMMARY OF THE INVENTION

Therefore, it is the object of the invention to propose a method and atleast one device by which it is possible to position and align samplesin the detection area of a microscope arrangement with high efficiency.By the word positioning is meant within the meaning of the invention thearrangement of the sample in the detection area of the objective, whilethe word alignment refers to the respective viewing direction of theobjective on the sample.

According to the invention, the following method steps are carried outin a method of the type mentioned above:

-   -   the sample is introduced into a transparent medium which is        initially in liquid state,    -   the medium is changed from the liquid state to a solid state so        that the sample is fixated inside the medium, but the medium        remains transparent or the transparency changes only negligibly,    -   the solidified medium is positioned in the microscope        arrangement in such a way that the sample enclosed therein is        situated in the detection area of the objective.

In a first preferred embodiment of the invention, a plurality of samplesare introduced into the transparent medium which is initially still in aliquid state. The medium with the samples contained therein is stored ina sample reservoir.

When one of the samples is to be examined, a partial amount of thetransparent medium which is still in liquid state is removed from thesample reservoir with the sample to be examined. After being removed,the partial amount of medium is changed from liquid to solid state, andthe sample is fixated within the solidifying partial amount of medium.

A partial amount can also be removed with a plurality of selectedsamples. After removal, this partial amount of the medium is changedfrom the liquid state to the solid state, and the samples are fixatedwithin the solidified partial amount of medium.

The partial amount of medium is removed from the sample reservoir, forexample, in that it is sucked into the hollow space of a capillary orcannula or, for example, into the hollow space of a disposable medicalsyringe.

By capillary is meant within the meaning of the present invention ahollow needle with a very small inner diameter and a suction anddelivery piston which is displaceably guided in the interior; a suctioneffect is achieved when the suction and delivery piston is displaced inone direction so that a partial amount of the liquid medium is removedfrom the reservoir, while the medium with the enclosed sample is pushedout of the capillary when the suction and delivery piston is displacedin the opposite direction.

By cannula is meant within the meaning of the invention a hollow needlewith an inner diameter greater than that of the capillary and,consequently, without capillary action. A displaceable suction anddelivery piston can likewise be provided in the interior of the cannulafor the purpose described above.

A suction and delivery piston which can be used to suck in or push out apartial amount of medium is likewise provided in the, usuallycylindrical, hollow space of the disposable syringe.

In a method step following the fixating of the sample, the solidifiedmedium is held in the microscope arrangement in such a way that thesample fixated therein is situated in the detection area. For thispurpose, at least that portion of the solidified medium in which asample is located is pushed out of the hollow space into the detectionarea. The portion of medium that is not pushed out remains in the hollowspace and is held therein, and the alignment and position of the samplein the detection area is influenced in a desired manner by means ofspecific changes in the position of the capillary, cannula or disposablesyringe.

If there are a plurality of enclosed samples, the medium is held in themicroscope arrangement in such a way that one of the samples enclosedtherein is initially situated in the detection area, and after thissample has been examined the next sample located in the partial amountis positioned and aligned and can be examined.

In a second preferred embodiment of the invention, the transparentmedium which is still in the liquid state is stored in a mediumreservoir without samples.

In contrast to the first embodiment of the invention:

-   -   a partial amount of the medium which does not yet contain a        sample is initially introduced into the hollow space of a        capillary, a cannula or a disposable syringe in a first step,        and    -   one or more samples are introduced into the medium already        located in the hollow space in a second step.

The inventive idea includes different ways of carrying out the method:For example, it is conceivable to carry out both of the above-mentionedsteps directly one after the other in that the medium in the liquidstate is introduced first and, immediately following this, the sample isintroduced into the medium which is still in the liquid state. This canbe carried out with just a sample by itself or also with a sample whichis already embedded in a smaller partial amount of the medium.

When a disposable syringe is used, for example, the liquid medium can beintroduced first and then one or more samples can be introduced into thecylindrical hollow space of the disposable syringe making use ofgravitational force.

In contrast to this, it can also be provided that:

-   -   the medium introduced in the hollow space is first changed from        the liquid state to the solid state in an intermediate step        before introducing the samples, or    -   a partial amount of the medium which is in the solid state is        introduced into the hollow space, and    -   the medium in the hollow space is not liquefied again until a        later time,    -   the sample is introduced, and    -   the medium is solidified again in order to fixate the sample        therein.

This way of carrying out the method can be used advantageously inconnection with preparing a plurality of samples which are alreadyfixated prior to microscopic examination.

In each of the cases mentioned above, the solidified medium is pushedout of the hollow space in order to position and align a sample fixatedtherein in the detection area of the objective of a microscopearrangement.

The change of the medium from the liquid state to the solid state, orvice versa, is carried out under external influences, particularly byheating or cooling. The solidification of the medium can also be carriedout under the influence of light.

Further, an automation of the method steps in their entirety or anautomation of some of the individual method steps lies within the scopeof the invention.

A curable gel, preferably agarose gel, is used as medium.

The invention is further directed to a device for positioning andaligning a preferably biological sample in the detection area of anobjective of a microscope arrangement comprising:

-   -   a reservoir for a transparent medium which is initially still        liquid,    -   means designed for        -   removing a partial amount of the liquid medium and for            introducing a sample in this partial amount, or        -   introducing samples into the liquid medium inside the            reservoir and removing a partial amount of the medium with a            sample contained therein,    -   means for changing the removed partial amount of medium from the        liquid state to the solid state, at least one sample being        fixated within the partial amount of the medium, and    -   a device for positioning and aligning the solidified partial        amount of medium in the microscope arrangement in such a way        that a sample contained therein is situated in the detection        area of the objective.

The device is advantageously outfitted with a manipulating unit whichhas a capillary, a cannula or a disposable medical syringe in which asuction and delivery piston is movably guided. Small volumes can besucked in by displacing the piston and by the vacuum pressure generatedin this way. Accordingly, it is possible to remove a partial amount onthis order of magnitude from the total reservoir of still liquid mediumin a precise manner.

The manipulating unit will be explained more fully referring to theexample of capillaries, although the invention is not limited to this.

For example, the manipulating unit can be designed to receive aplurality of capillaries simultaneously. This proves advantageous when aset of samples is prepared and distributed to a plurality ofcapillaries, which samples are received by the manipulating unit andexchanged with one another in a simple manner, so that the samplesreceived therein can be examined efficiently one after the other inrapid sequence.

For the purpose of exchanging the capillaries in the sample space, themanipulating unit can be outfitted with a turret arrangement receivingthe, capillaries. Accordingly, one of the capillaries is moved into aposition in which the suction and delivery piston is grasped and theportion of the medium with the enclosed sample is pushed into thedetection area by means of the suction and delivery piston. After beingexamined, the sample is pulled back into the capillary by the suctionand delivery piston and is stored therein, the next capillary with thenext sample is moved into position, the suction and delivery piston isgrasped, and this sample is now pushed into the detection area.

In this connection, it is advantageous when the suction and deliverypiston comes into contact with the medium directly (i.e., without an aircushion) so as to facilitate metering of a highly viscous medium such asa gel in particular. Further, this prevents unwanted substances beingsucked in via the air cushion (e.g., in the form of aerosols) which isimportant above all when handling living biological samples.

In a particularly advantageous embodiment of the arrangement accordingto the invention, the capillary, including the suction and deliverypiston guided therein, is connected to the manipulating unit byconnection elements which can be disconnected manually. In so doing, thecapillary can also be located in a sleeve which is then connected to themanipulating unit.

In this way, it is possible to disconnect the capillary, including thesuction and delivery piston guided therein, from the manipulating unit,to immerse the suction opening of capillary in the medium, to displacethe suction and delivery piston in the capillary, and to suck a partialamount of medium into the capillary corresponding to a predeterminedvolume.

Depending on one of the modes of carrying out the method which havealready been described, the sucked in medium can already contain asample, or a sample is introduced subsequently in the partial amount ofmedium located in the capillary.

When the medium and the sample are located in the capillary, the mediumis changed from the liquid state to the solid state with the meansaccording to the invention provided for this purpose in order to fixatethe sample within the partial amount of medium as will be described inmore detail below with reference to an embodiment example.

The capillary which now contains the solid medium and the sampleenclosed therein is subsequently arranged at the manipulating unit againby means of the connection elements.

The manipulating unit, per se, is fastened to the microscope arrangement(e.g., by means of a straight-line guide) and it is outfitted with meansfor changing the position and alignment of the capillary relative to themicroscope arrangement, wherein there is a change in position of thecapillary in coordinates X, Y, Z and a rotational movement around thelongitudinal direction of the capillary by an angle φ.

With the manipulating unit it is possible to move the end portion of thecapillary in which the sample is located into the vicinity of thedetection area initially by means of displacement in coordinates X and Yand then, using the suction and delivery piston, to push the solidifiedmedium with the enclosed sample out of the capillary in direction ofcoordinate Z until the sample is positioned in the detection area. Byrotating the sample around angle φ, the alignment of the sample (i.e.,the viewing direction of the objective on the sample positioned in thedetection area) is varied in the desired manner.

It lies within the scope of the invention to directly manually initiatethe movements of the capillary by means of correspondingly constructedgear unit members and also to initiate these movements by controllingmotorized drives to which the capillary is connected by gear unitmembers.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a capillary with a suction and delivery piston guided so asto be movable in its interior and a sample reservoir in which the endportion of the capillary is immersed;

FIG. 2 shows the capillary according to FIG. 1 in a curing station,shown schematically, in which heat energy is extracted from atransparent medium which is initially still in liquid state in order tosolidify it;

FIG. 3 shows a schematic illustration of the mode of operation of amanipulating unit designed for positioning and aligning a sample in thedetection area of a microscope objective;

FIG. 4 shows the manipulating unit according to FIG. 3, wherein thesample is positioned in the illumination beam path of a microscope andis aligned on the sample in a first viewing direction of the microscopeobjective on the sample;

FIG. 5 shows an alternative example for introducing samples into acapillary shown in FIG. 1;

FIGS. 6 to 8 show another example for introducing individual samplesinto a curable medium; and

FIG. 9 shows an example for a timed introduction of a plurality ofsamples into a curable medium.

DETAILED DESCRIPTION OF EMBODIMENTS

It is to be understood that the figures and descriptions of the presentinvention have been simplified to illustrate elements that are relevantfor a clear understanding of the present invention, while eliminating,for purposes of clarity, many other elements which are conventional inthis art. Those of ordinary skill in the art will recognize that otherelements are desirable for implementing the present invention. However,because such elements are well known in the art, and because they do notfacilitate a better understanding of the present invention, a discussionof such elements is not provided herein.

The present invention will now be described in detail on the basis ofexemplary embodiments.

FIG. 1 shows a capillary 1 with a suction and delivery piston 2 which isguided in the interior so as to be displaceable in directions R1 and R2.Combinations of capillaries 1 and suction and delivery pistons 2 of thiskind are known, per se, as pipettes and are used for dispensing liquids.

The capillary 1 can be made of glass or plastic and can be provided witha volume scale (not shown in the drawing) arranged laterally inlongitudinal direction. The suction and delivery piston 2 is generallymade of a flexible plastic, but can also be formed of a stainless steelrod linkage with a plunger arranged thereon.

When the end portion 3 of the capillary 1 is dipped into a samplereservoir 4 in which a transparent, initially liquid medium in the formof agarose gel 5 and a plurality of samples 6 are located and thesuction and delivery piston 2 is displaced inside the capillary 1 indirection R1, a partial amount of the agarose gel 5 of for example,about 30 μL to 50 μL is sucked into the capillary 1 and this partialamount of agarose gel 5, including one of the samples 6, is removed fromthe total reservoir of samples 6 in a precise manner.

The sample 6, surrounded by the agarose gel 5 which is likewise removed,is transported by the capillary 1 to a curing station 7, shownschematically in FIG. 2, where the agarose gel 5 is cooled. The agarosegel 5 is increasingly solidified as heat energy is extracted whileremaining transparent, and the sample 6 is fixated in the agarose gel 5.

The capillary 1 with the sample 6 fixated in the agarose gel 5 is nowtransported to a manipulating unit 8, shown schematically in FIG. 3, andfixed therein by means of a receptacle 9. The manipulating unit 8 is inturn fastened to a microscope stand 10, only a partial area of which isshown for the sake of clarity.

It is advantageous when a connection of the manipulating unit 8 to themicroscope stand 10 is provided by means of straight-line guides whichensure a displacement of the manipulating unit 8 relative to themicroscope stand 10 in coordinates X and Y

The manipulating unit 8 is outfitted with an actuating element 11 whichis supported in a rotating and straight-line guide 12 and is accordinglydisplaceable in directions R1 and R2 and rotatable around an angle φ.The directions R1 and R2 extend parallel to coordinate Z in coordinatesystem X, Y, Z.

The actuating element 11 has a clamping device 13 which encloses the endof the suction and delivery piston 2 remote of the sample 6. Theclamping device 13 causes displacements of the actuating element 11 indirections R1 and R2 and also the rotation of the actuating element 11to be transmitted to the suction and delivery piston 2. A drive element14 serves to initiate the displacements in directions R1 and R2 and therotational movement.

After the capillary 1 is locked in the manipulating unit 8 and theclamping connection between the suction and delivery piston 2 and theactuating element 11 is produced, the sample 6 is located in thevicinity of the detection area which is represented here by theillumination beam path 15 in the form of a light sheet which is formedand provided for subsequent examination of the sample 6 by the method ofsingle plane illumination microscopy (SPIM).

Before starting the examination, it must be ensured that the sample 6 issituated in the illumination beam path 15. To achieve the configurationshown in FIG. 4, the actuating element 11 is displaced in direction R2by the rotational movement of the drive element 14 based on the diagramshown in FIG. 3, and the displacing movement is transmitted by means ofthe clamping device 13 to the suction and delivery piston 2 and then tothe agarose gel 5 with the sample 6 enclosed therein.

Since the capillary 1 is not included in this displacing movementbecause it is locked in the manipulating unit 8, the agarose gel 5 withthe sample 6 is pushed out of the capillary 1 until the configurationillustrated in FIG. 4 is achieved and the sample 6 is situated in theillumination beam path 15.

The detection direction of the microscope objective, not shown in thedrawing, is perpendicular to the drawing plane. By rotating thecapillary 1 by an angle φ within a range of 360 degrees, the detectiondirection relative to the sample 6 can be changed as needed.

In this way, the positioning and alignment of samples 6 in theillumination beam path 15 and detection area of the microscope,respectively, can always be reproduced.

A first variant for filling the capillary 1 was described with referenceto FIG. 1. An alternative variant which satisfies the demand forincreased throughput per time unit in the examination of samples 6 isshown by way of example in FIG. 5.

In this case, a filling station 16 is provided in which an emptycapillary 1 is initially inserted. The filling station has an access 17for agarose gel 5 and an access 18 for samples 6. Further, a guide 19 isprovided for a piston 22 in order to displace the latter in a straightline in directions R1 and R2. The filling station is preferably combinedwith a curing station possessing possibilities for temperature controland for supplying and removing heat.

Valves 20 and 21 which are preferably electronically controllable andare alternately opened and closed depending on the control are arrangedin accesses 17 and 18.

The filling station 16 is operated in such a way, for example, that thevalve 20 is initially open and liquid agarose gel 5 is displaced throughthe access 17 until it is below the piston 22 and is displaced fartherinto the capillary in direction R2.

The agarose gel 5 is pressed in direction R2 into the capillary 1 orsinks (for example, when the piston 22 is removed) into the capillary 1under the influence of gravitational force or capillary force. Toprevent the agarose gel 5 from flowing out through the lower end of thecapillary 1, a closure 23 is placed on this end as soon as agarose gel 5is located in the capillary 1. The valve 20 is then closed.

Valve 21 is now opened and a sample 6 is displaced through access 18until it is below the piston 22 and is displaced farther into thecapillary in direction R2.

After valve 21 is closed, the valve 20 is opened again, if required, andliquid agarose gel 5 is again fed through access 17.

Alternatively, instead of supplying a sample 6 by itself, a sample 6which is already embedded in a partial amount of agarose gel 5 can befed through access 18. This partial amount is then pressed into thecapillary 1 along with the embedded sample 6 by means of the piston 22or sinks into the capillary 1 under the influence of gravitational forceand combines with the agarose gel 5 already located therein.

Subsequently, the valves are closed and the piston 2 is displaced indirection R2 so that it contacts the agarose. After the agarose cures,the sample can be moved up and down by rotating the drive element 14.

It is advantageous when the upper opening of the capillary 1 (i.e., theopening of the capillary 1 opposite the direction of gravitationalforce) is conically expanded so that the agarose gel 5 can flow into thecapillary 1 more reliably (not shown in the drawing).

It also lies within the scope of the invention to construct the fillingstation in the manner shown in FIG. 6. Filling with agarose gel 5 isaccordingly initially carried out as described above. However, therounded or otherwise shaped end of a tool guided through access 18 isthen pressed into the agarose gel 5 which is still in liquid state,whereupon the agarose gel 5 is cured, and the tool is removed againafter curing so that a depression 24 (e.g., in the shape of a hollowcone or a trough) remains in the cured agarose gel 5. The valves are notshown in FIG. 6 for the sake of clarity, especially since their functionhas already been described referring to FIG. 5.

After the tool has been removed, a sample 6 is advanced into thedepression 24 through access 18 as is indicated in FIG. 7. Just thesample by itself or the sample located in a partial amount of agarosecan be supplied. If required, an additional partial amount of agarose isintroduced.

When the sample 6 has been advanced into the depression 24, the agarosegel 5, including the sample 6 located in the depression 24, is advancedby the piston 2 until the configuration shown in FIG. 8 is achieved. Ifrequired, the agarose gel 5 is now liquefied again by temporarilysupplying heat in order to embed the sample 6 completely in the agarosegel 5.

It is advantageous when the filling station is designed so as to becompatible with a microscope so that the sample can be inserted in thedepression 24 and oriented while being observed.

Subsequently, the same process as that described referring to FIG. 5 maybe carried out, wherein the capillary 1 is removed from the fillingstation 16 and is prepared for microscopic examination as was describedabove with reference to FIG. 3 and FIG. 4.

It is advantageous when the filling station and the manipulating unitform a functional unit.

However, the inventive idea also includes a mode of operation forfilling capillaries or cannulas which expands on the mode of operationdescribed with reference to FIGS. 6 to 8. The following descriptionrefers to FIG. 9.

The following is carried out at given time intervals:

-   -   a first sample 6.1 is introduced into the agarose gel 5,    -   the agarose gel 5 is advanced with the piston 22,    -   a second sample 6.2 is introduced into the agarose gel 5,    -   the agarose gel 5 is advanced again,    -   a third sample 6.3 is introduced, and so on, until a given        quantity n of samples 6.n have been inserted in the agarose gel        5, wherein distances a are adjusted between the samples 6.1,        6.2, . . . , 6.n depending on the timing and forward feed speed.

In this case, the agarose gel 5 is solidified by extracting heat atposition P with the same timing with which the samples 6.1, 6.2, . . . ,6.n are embedded in the agarose gel 5, and the sample 6.2 located atposition P is accordingly fixated in the agarose gel 5.

The samples 6 can subsequently be examined microscopically as wasdescribed above.

A portion A of the agarose gel 5 in which a sample (e.g., sample 6.1 inthis instance)is embedded is severed by means of a cutting device 26which is guided through the housing wall of the curing station 25 andprovided with a knife 27 which is displaceable in directions S1 and S2.

The severed portion falls through a funnel-shaped opening 28 out of thecuring station 25 under the influence of gravitational force and can besupplied for further analytic methods.

It is noted that the suction and delivery piston 2 and the piston 22described in the embodiment examples have different functions inasmuchas the piston 22 has no suction function, which is achieved, forexample, in that it is guided with a sufficiently large play in thehollow cylinder. Nevertheless, the piston 22 can be exchanged for asuction and delivery piston 2 insofar as the corresponding function isdesired for handling the agarose gel 5 and sample 6.

While this invention has been described in conjunction with the specificembodiments outlined above, it is evident that many alternatives,modifications, and variations will be apparent to those skilled in theart. Accordingly, the preferred embodiments of the invention as setforth above are intended to be illustrative, not limiting. Variouschanges may be made without departing from the spirit and scope of theinventions as defined in the following claims.

REFERENCE NUMBERS

-   1 capillary-   2 suction and delivery piston-   3 end portion-   4 sample reservoir-   5 agarose gel-   6 sample-   7 curing station-   8 manipulating unit-   9 receptacle-   10 microscope stand-   11 actuating element-   12 rotating and straight-line guide-   13 clamping device-   14 drive element-   15 illumination beam path-   16 filling station-   17, 18 access-   19 straight-line guide-   20, 21 valve-   22 piston-   23 closure-   24 depression-   25 curing station-   26 cutting device-   27 knife-   28 opening-   R1, R2 directions-   φ angle-   a distance-   A portion-   P position

1. A method for positioning a sample in the detection area of anobjective in a microscope arrangement having the following method steps:introducing a sample into an initially liquid, transparent medium;changing the medium from the liquid state to the solid state so that thesample is fixated inside the medium, but the medium remainssubstantially transparent; and positioning the solidified medium in themicroscope arrangement in such a way that the sample enclosed therein issituated in the detection area of the objective.
 2. The method accordingto claim 1, further comprising: introducing one or more samples into thetransparent medium which is initially liquid; storing the medium withthe samples in a sample reservoir; removing a partial amount of themedium with a sample contained therein from the sample reservoir; afterthe removing step, changing the partial amount of medium from the liquidstate to the solid state, wherein the sample is fixated within thepartial amount of medium; and positioning the cured partial amount ofmedium in the microscope arrangement in such a way that the samplecontained therein is situated in the detection area of the objective. 3.The method according to claim 1; wherein the partial amount of mediumwhich is still in the liquid state with the sample contained therein issucked into the hollow space of a capillary, cannula or disposablesyringe; wherein the medium inside the hollow space is then changed fromthe liquid state to the solid state; and wherein the solid medium withthe enclosed sample is ejected from the hollow space.
 4. The methodaccording to claim 1, further comprising: storing the transparent mediumwhich is initially liquid on a medium reservoir; removing a partialamount of the medium from the medium reservoir; introducing a sampleinto the partial amount of medium which is still liquid; after theremoving step, changing the partial amount of medium from the liquidstate to the solid state, wherein the sample (6) is fixated within thepartial amount of medium; and positioning the cured partial amount ofmedium in the microscope arrangement in such a way that the samplecontained therein is situated in the detection area of the objective. 5.The method according to claim 4; wherein the partial amount of mediumwhich is still liquid is introduced into the hollow space of acapillary, a cannula or a disposable syringe; wherein the partial amountof medium which is still liquid is held inside the hollow space; andwherein a sample is introduced into the partial amount located in thehollow space.
 6. The method according to claim 5; wherein one of the twoend openings of the hollow space containing the medium which is still inthe liquid state is hermetically closed; and wherein the method furthercomprises: introducing the sample into the medium which is still in theliquid state inside the hollow space through the opposite second endopening; next, changing the medium inside the hollow space from theliquid state to the solid state; and next, ejecting the solid mediumwith the enclosed sample (6) is ejected from the hollow space.
 7. Themethod according to claim 1; wherein the change of the partial amountfrom the liquid state to the solid state is carried out under externalinfluences on the medium.
 8. The method according to claim 1; wherein acurable gel is used as medium.
 9. The method according to claim 1;wherein an automation of the method steps in their entirety or anautomation of some of the individual method steps is provided.
 10. Themethod according to claim 9; wherein a first sample is introduced intothe gel in the hollow space at predetermined time intervals; wherein thegel is pushed forward; wherein a second sample is introduced into theagarose gel; wherein the gel is pushed forward again; and wherein athird sample is introduced into the gel, and so on, until a givenquantity n of samples has been introduced into the gel, whereindistances a are adjusted between the samples depending on the timing andforward feed speed.
 11. A device for introducing a sample into thedetection area of an objective of a microscope arrangement, comprising:a reservoir for a transparent medium which is initially still liquid;means designed for: removing a partial amount of the liquid medium andfor introducing a sample into this partial amount; or introducingsamples into the liquid medium inside the reservoir and removing apartial amount of the medium with a sample contained therein; means forchanging the removed partial amount of medium from the liquid state tothe solid state, wherein at least one sample is fixated within thepartial amount of the medium; and a device for positioning and aligningthe solidified partial amount of medium in the microscope arrangement insuch a way that the sample contained therein is situated in thedetection area of the objective.
 12. The device according to claim 11,further comprising: a manipulating unit which has a hollow needle inwhich a suction and delivery piston is movably guided for sucking apartial amount of the medium into the hollow needle or ejecting it fromthe hollow needle.
 13. The device according to claim 12; wherein themanipulating unit is connected to the microscope arrangement by aholder, and the holder is constructed with means for changing theposition and alignment of a capillary or cannula relative to themicroscope arrangement, wherein there is a change in position incoordinates X, Y, Z and a rotational movement around the longitudinaldirection of the capillary or cannula by an angle φ.
 14. The deviceaccording to claim 11; wherein a capillary, including the suction anddelivery piston, is fastened to the manipulating unit by means ofconnection elements which can be disconnected manually.
 15. A device forembedding a plurality of samples in a curable transparent medium,comprising. an access for the medium, an access for the samples; forwardfeed devices for the medium and the samples; a device for introducingthe samples individually into the medium successively in a timed manneruntil a given quantity n of samples has been inserted into the medium;and wherein the medium is advanced in a timed manner, and distances aare adjusted between the samples within the medium depending on thetiming and forward feed speed.
 16. The device according to claim 12;wherein the hollow needle of the manipulating unit is in the form of acapillary or a cannula.